WO2016113473A1

WO2016113473A1 - O-quinone compounds as agents neutralising nitric oxide

Info

Publication number: WO2016113473A1
Application number: PCT/FR2015/050064
Authority: WO
Inventors: Daniel Jean; Maryse Pouligon
Original assignee: Institut des Substances Végétales
Priority date: 2015-01-12
Filing date: 2015-01-12
Publication date: 2016-07-21
Also published as: ES2797056T3; CA2973047A1; US20170369414A1; EP3244882A1; EP3244882B1

Abstract

The invention relates to O-quinone compounds of general formula (I) as agents neutralising nitric oxide, and to the therapeutic or cosmetic use thereof.

Description

O-Quinone Compounds as Nitric Oxide Neutralizers

The present invention relates to the field of the treatment of diseases and / or inflammatory disorders resulting from an excess of nitric oxide (NO); it relates more particularly to o-quinonic compounds as neutralizing agents nitric oxide and their therapeutic or cosmetic use.

Nitric Oxide (NO) is one of the smallest molecules produced by biological systems. Its actions are as diverse in organisms as the cells capable of producing it. Indeed, its relatively unstable free radical status that gives it its electronic structure (its half-life is 5 seconds) allows it to act on a large number of molecules more or less sensitive to its action within cellular metabolic processes and extracellular.

It was shown in the 1980s that nitric oxide acts specifically as a mediator of vascular dynamics by driving the relaxation of vessels. Amongst other cells, during inflammatory processes, this molecule is produced by endothelial cells (1,2) and macrophages (3,4) by enzymes, nitric oxide synthases (NOs), which catalyze the transformation of arginine. in citrulline. These enzymes exist constitutionally or are induced during immune reactions, this is called iNOs. Vasodilation is induced by stimulation of smooth muscle guanylate cyclase (5). Nitric oxide is also formed by endothelial cells in response to a variety of substances such as bradykinin (2), histamine and 5-hydroxytryptamine (6). Nitric oxide can also be formed by macrophages activated by lipopolysaccharides or cytokines (6).

Nitric oxide also plays a role in immunity thanks to its high chemical reactivity which allows it to participate in the lysis of phagocytosed seeds by macrophages. In the presence of the super-oxide radical, nitric oxide is converted into peroxynitrite, itself a particularly aggressive agent as oxidant.

The induction of nitric oxide synthesis has been shown during the inflammatory response initiated by microbial products during infections or autoimmune reactions. Nitric oxide is therefore a mediator of physiology that has a positive side by regulating vasodilatation and participating in immune function, and a negative side when it is emitted in excessive amounts. In excess, it can produce inflammation, destruction of cells and tissues and can also cause vasodilation leading to pain or fatal hypotension in septic or anaphylactic shock.

It has been shown that nitric oxide has an influence on the proliferation of keratinocytes (7) and very high levels of plasma nitric oxide have been observed in subjects with psoriasis (8), suggesting the influence of mediator on this disease.

Atopy is a predisposition to the amplified reaction of the immune response. In its dermatological form, atopic dermatitis affects about 15% of children in developed countries. High nitric oxide production has been shown to be involved in inflammation, vasodilation and oxidative damage to cells and skin tissues of subjects with atopic dermatitis (9).

Nitric oxide is also involved in solar erythema (10).

Similarly, at the joint level, nitric oxide is involved in the inflammatory manifestations of arthritis. It has also been shown in animal models that administration of NOs inhibitors significantly reduces cartilage inflammation (22).

Regarding the systemic inflammatory response to which septic shock refers, it produces generalized vasorelaxation due to the massive production of nitric oxide, which results in hypotension that can lead to death in the absence of appropriate treatment applied in emergency ( 11). This mechanism is also that which develops during anaphylactic shock (12).

Consequently, it may sometimes be desirable to supply nitric oxide to an organ that lacks it, as is the case, for example, in chronic hypertension associated with myocardial infarction suites or to prevent it. in many other cases it would be necessary to neutralize the same nitric oxide to reduce its harmful effects, either in an ambulatory manner as in atopic dermatitis, eczema, psoriasis, cough, or osteoarthritis, or massively urgent way to treat collapse associated with septic or anaphylactic shock.

Nitric oxide production can be reduced by inhibiting nitric oxide synthases (NOs), enzymes that release it from L-arginine, or by chemically neutralizing nitric oxide or by capturing it with appropriate complexing agents. . Inhibitors of NOs, among them arginine derivatives, such as the methylated, nitrated or propylated derivatives of this amino acid (13), are known, but their use in systemic therapy has serious drawbacks (14). Indeed, the inhibition of NOs is difficult to control and to avoid an overdose that can lead to pulmonary hypertension, it is often necessary to administer nitric oxide gas through the respiratory tract at the same time as the NO inhibitor , while monitoring the cardiovascular parameters of the patient. This disadvantage could be due to the lack of specificity of the inhibitors used vis-à-vis the different isoforms of the NOs, but for the moment, no specific inhibitor has proved its effectiveness and its reliability in clinical studies.

NOs inhibitors have been proposed to treat aesthetic disorders of the skin (15, 16) but their use is limited by the harmful side effects, such as arterial hypertension, that they can manifest and the fact that these inhibitors are all synthetic products, whereas today consumers prefer to turn to substances of natural origin.

There are also molecules capable of neutralizing nitric oxide, such as hemoglobin, which is considered the reference molecule in this field, and various synthetic molecules such as carboxy-2-phenyl-4,4,5,5- tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO) (17) and ruthenium derivatives (18). The use of all these molecules has drawbacks in particular for therapeutic and cosmetic applications; for example, ruthenium derivatives are considered potentially toxic and carcinogenic.

Apart from these three groups of molecules, a very large number of other components of plant origin have been proposed as possible neutralizing agents for nitric oxide, such as flavonoids and more generally polyphenols. But the evaluation of their neutralizing effect by the Griess method, which measures the N0 ₂ - ion, does not make it possible to evaluate the real effect of the molecule tested. Indeed, any antioxidant capable of lowering the concentration of oxygen radicals in the medium is an inhibitor of the reaction that leads from NO to the N0 - ion, and thus the Griess method provides a misleading indication of the actual effect of the molecule on its neutralizing power of nitric oxide, because it is impossible to distinguish between the antioxidant power and the neutralizing power vis-à-vis the nitric oxide. In order to have a real appreciation of the neutralizing effect of the compound with respect to nitric oxide, the inventors have developed a specific in vitro method which directly evaluates the concentration of nitric oxide in the medium and who can measure the neutralization in real time in the presence of the molecules tested; Example I describes this method.

For this, the nitric oxide is prepared in a separate container by performing the reduction of sodium nitrite by ferrous sulfate and driven by a stream of nitrogen gas in another container containing the test product in solution in a buffer. In this solution is placed an amperometric probe which measures in real time the current produced by the presence of nitric oxide in solution.

Once provided with this specific method of evaluating the nitric oxide neutralization, the inventors were able to confirm the neutralizing effect of certain molecules known as such, such as hemoglobin, but surprisingly also discovered that certain molecules, as described as good nitric oxide neutralizers were not in reality, once screened by this specific method, including flavonoids such as quercetin, catechin and luteolin.

They have also identified molecules that are very good nitric oxide neutralizers and have never been described as such.

Thus, the present invention aims at the identification of effectively neutralizing compounds nitric oxide, easily used in therapy or in cosmetics and does not have the disadvantages of those described above.

The invention relates to a compound of formula (I):

wherein R ¹ is selected from the group consisting of:

• -CH = CH-COOR ²

with R ^{2 being} able to represent:

- H, an alkyl chain in C _r C ₃ linear, optionally substituted by one or more functions chosen from hydroxyl and carboxylic acid, a benzene ring, with a dihydric phenol or by a radical caffeoyl; A saturated or unsaturated C ₆ ring, optionally substituted with one or more functions chosen from hydroxyl and carboxylic acid functions and / or with a caffeoyl radical;

The formula -Y-O-X- (3,4) diphenol (II) with:

. Y is a pyranose ring, substituted with -CH ₂ -OH and at least one hydroxyl function and optionally substituted with rhamnose;

. X may represent the chain -CH ₂ -CH ₂ - or -CH ₂ -CH (OH) -;

• -ZOR ³

With:

Z may represent the chain -C¾-CH ₂ -, -CH (OH) -CH ₂ - or -CH ₂ -CH (COOH) -;

R ³ may represent:

. - CO-CH = CH- (3,4) diphenol or

. The formula -W-O-CO-CH = CH- (3,4) diphenol (III) with W being a pyranose ring, substituted with -CH ₂ -OH and at least one hydroxyl function and optionally substituted with a rhamnose;

for its use for the prevention and / or treatment of diseases and / or disorders resulting from an excess of NO.

By physiologically acceptable is meant compatible with administration to a subject, preferably a mammal, by any route of administration.

The excess of NO can be determined by measuring the level of nitrates in the blood serum (9). A human individual is then considered to have an excess of NO when his concentration of nitrates in the blood serum is greater than or equal to 14 mmol / L, in particular 30 mmol / L or 50 mmol / L.

According to a variant of the invention, the compound of formula (I) is such that R ¹ is selected from the group comprising:

• -CH = CH-COOR ²

with R ^{2 being} able to represent:

- H or

The formula -Y-O-X- (3,4) diphenol (II) with:

. Y represents a pyranose ring, substituted with -CH ₂ -OH and at least one hydroxyl function and substituted by rhamnose;

. X represents the chain -CH ₂ -CH ₂ -; • -ZOR ³

With:

Z represents the chain -CH ₂ -CH ₂ -;

- R ³ represents the formula -W-O-CO-CH = CH- (3,4) -diphenol (III) with W represents a pyranose ring, substituted with -CH-OH and at least one hydroxyl function and substituted by a rhamnose.

According to another variant of the invention, this relates to a compound of formula (Ia) such that:

with R ^{2 being} able to represent:

H, a linear C ₁ -C ₃ alkyl chain, optionally substituted with one or more functional groups chosen from hydroxyl and carboxylic acid functions, with a benzene ring, with a diphenol or with a caffeoyl radical; A saturated or unsaturated C ₆ ring, optionally substituted with one or more functions chosen from hydroxyl and carboxylic acid functions and / or with a caffeoyl radical;

The formula -Y-O-X- (3,4) diphenol (II) with:

. X can represent the chain -CH ₂ -CH ₂ - or -CH ₂ -CH (OH) -.

According to still other variants of the invention, this relates to a compound of formula (I) such that:

- R ¹ is -CH = CH-COOR ² and R ² is an H or a linear C 1 or C 2 alkyl chain; or

- R ¹ is -CH = CH-COOR ² and R ² corresponds to the formula -Y-O-X- (3,4) diphenol (II) with Y is the glucose substituted by a rhamnose and X can represent the chain - CH ₂ -CH ₂ - or -

CH ₂ -CH (OH) -; or

- R ¹ is -ZOR ³ with Z being able to represent the chain -CH ₂ -CH ₂ -, -CH (OH) -CH ₂ - or - CH ₂ -CH (COOH) -; and R ³ represents the formula -W-O-CO-CH = CH- (3,4) diphenol (III) with W is a glucose substituted with a rhamnose. According to a preferred variant of the invention, this relates to the compounds mentioned below or to their physiologically acceptable salts for their use for the prevention and / or the treatment of diseases and / or disorders resulting from an excess of NO:

The quinone of caffeic acid:

Quinone verbascoside, present in 2 forms:

First form of the quinone of the second form of the verbascoside verbascoside quinone

The term "optionally substituted linear C 1 -C 3 alkyl chain" denotes a saturated, linear, optionally substituted hydrocarbon-based chain of 1 to 3 carbon atoms, such as methyl, ethyl or propyl.

By hydroxyl function is meant the -OH group.

By carboxylic function, denotes the -COOH group.

By benzene ring denotes the aromatic functional group of formula C ₆ H ₆ .

By diphenol means an aromatic compound, consisting of a benzene ring and two hydroxyl functions, of empirical formula C ₆ H ₆ 0 ₂ .

By radical Caffeoyl, denotes the radical derived from caffeic acid, of formula: -O-CO-CH = CH- (3,4) diphenol. By pyranose ring is meant a saturated C ₆ ring consisting of 5 carbon atoms and one oxygen atom.

By way of example, mention may be made of glucose which has a pyranose ring, substituted by a -CH ₂ -OH and 4 hydroxyl -OH.

By rhamnose, we denote the ose under its conformation D or L, under its

The compounds of formula (I) can be chemically synthesized or extracted from plants.

As an example, orthoquinone of caffeic acid which can be obtained crystallized, by oxidation of caffeic acid by Po-chloranil, according to a method described in the prior art (20, 21).

Alternatively, the orthoquinone of caffeic acid can be extracted from plants such as Salvia officinalis, Mentha spicata, Cinnamomum verum, Thymus vulgaris.

Those skilled in the art are aware of methods for extracting chemical compounds from plants. These methods include extraction with chemical solvents, with a supercritical fluid such as C0 ₂ , nanofiltration.

Quinone of verbascoside can be extracted from plants of the Lamiaceae family (Phlomis, Scrophulariaceae, Verbascum phlomoides, Verbascum mallophorum), or the Buddlejaceae family (Buddleja globosa, Buddleja cordata), or the family Bignoniaceae (Pithecoctenium). sp, Tynanthus panurensis), or hungry Orobanchaceae (Cistanche sp, Orobanche rapum-genistae), in the family Plantaginaceae (Plantago lanceolata, Verbenaceae, Verbena officinalis, Aloysia citrodora) in the family Oleaceae (Olea europaea) in the family Lentibulariaceae (Pinguicula lusitanica) and in the family Byblidaceae (Byblis liniflora). In addition, the inventors have demonstrated, surprisingly and contrary to what was supposed in the prior art (19), that an extract of a plant known for its anti-inflammatory effect, the plantain lanceolate {Plantago lanceolata), does not owe its anti-inflammatory effect to the phenylhydantoic glycoside, the verbascoside it contains, but to the quinone derived from this polyphenol.

Thus, according to another variant of the invention, it relates to an extract of Plantago lanceolata selectively enriched in quinone verbascoside; such an extract of Plantago lanceolata enriched in verbascoside quinone is preferably obtained from aerial parts of the plant by alcoholic maceration; such an extract can be obtained according to the process described in Example 5. This particular extract produces a significant effect of NO neutralization.

The invention relates more particularly to the use of a compound or extract of Plantago lanceolata according to the invention for the prevention and / or the treatment of inflammatory diseases and / or disorders.

In particular, the compounds or the Plantago lanceolata extract according to the invention are useful for the prevention and / or the treatment of diseases and / or inflammatory skin disorders such as psoriasis, atopic dermatitis, contact dermatitis, skin irritation, contact hypersensitivity reaction, skin allergic manifestations, excessive vasodilatation, rosacea, solar perythema, acne.

The compounds or plantago lanceolata extract according to the invention are still useful for the prevention and / or treatment of inflammatory diseases and / or articular disorders such as arthritis, including rheumatoid arthritis, infectious arthritis and posteoarthritis. osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, lupus erythematosus, chondritis.

The compounds or the extract of Plantago lanceolata according to the invention can be used to prevent or treat cough, whether it is associated with a respiratory tract disease such as bronchiolitis in infants, bronchitis, colds, whooping cough, tuberculosis, a bronchial cancer or associated with a particular state and / or environment as is the case of smoker's cough or caused by passive smoking, allergic cough, cough associated with asthma or gastro-oesophageal reflux.

The compounds or plantago lanceolata extract according to the invention can still be used to prevent or treat associated cardiovascular collapse. hypovolemic shock, or cardiogenic shock or anaphylactic shock or septic shock. Cardiovascular coUapsus corresponds to a collapse of blood pressure; the systolic blood pressure then becomes less than 80 mmHg.

The use of compounds or plantago lanceolata extract according to the invention may also have a cosmetic purpose, in order to fight against the intrinsic or extrinsic aging of the skin, more particularly to fight against the signs of skin aging, such as photoaging, wrinkles, fine lines, dry or cracked skin.

Among the other uses of these compounds or of the Plantago lanceolata extract according to the invention in cosmetics, mention may be made of:

- fight against neurogenic skin inflammatory processes;

improving the comfort of sensitive skin;

strengthening of the barrier function of the skin;

stimulating the hydration of the skin;

- improving the comfort of dry skin;

- control of sweating;

- stimulation of lipolysis;

- inhibition of hair loss.

The present invention also relates to a pharmaceutical or cosmetic composition containing at least one compound of formula (I) or an extract of Plantago lanceolata according to the invention and a physiologically acceptable vehicle.

Those skilled in the art will be able to adapt the formulation of the compositions according to the invention according to their physico-chemical properties and their route of administration.

The compositions according to the invention may be administered by any route of administration which notably includes the topical route, in particular on the skin or the hair, but also the oral route.

Preferably, when the compositions according to the invention are used for the treatment of diseases and / or inflammatory disorders of the skin or joints, or when they are used for a cosmetic purpose, they are administered by the topical route and may be chosen among a cream, a gel, an oil, a lotion, a milk. When the compositions according to the invention are used to treat cough, it will be preferred to administer them orally. The dosage form chosen will be a syrup, a lozenge to be sucked, a capsule, a tablet.

FIGURE:

Figure 1: Apparatus for implementing the specific method for evaluating the nitric oxide neutralization as described in Example 1.

EXAMPLES

Example 1. Evaluation Method for the Neutralization of Nitric Oxide

The nitric oxide is prepared extemporaneously in a container A, closed by a flexible polymer plug through which pass an incoming tubing and an outgoing tubing, with an internal diameter of between 50 μιη and 300 μη, but particularly of 100 μηι ( see FIG. 1), with a volume of between 10 ml and 100 ml, but particularly of 30 ml, by adding a volume of between 0.1 and 2 ml, but especially of 0.5 ml of an aqueous solution of sodium nitrite (NaNO ₂ , Sigma-Aldrich No. 237213) at a concentration of between 0.10% and 2.00%, but especially 0.70%, at a volume of between 5 ml and 80 ml of the solution following aqueous

Sulfuric acid (H ₂ S0 ₄ , Sigma-Aldrich reference 32,050-1): 1 ml

Ferrous sulphate (FeSO ₄ , 7H ₂ O, Sigma-Aldrich number 215422): 5.5 g

Deionized water: 200 ml

The container B in which the nitric oxide is measured is a light-opaque container, with a double jacket, allowing a circulation of water maintaining the temperature at 25 ° C., closed with a plug through which the tubing coming from the container passes. A and an exhaust manifold (see FIG. 1), with a volume of between 10 ml and 100 ml, preferably 30 ml, which receives a volume of between 5 ml and 80 ml, preferably 20 ml, of an aqueous solution of phosphate buffer pH 7.4.

Once mixed the solutions are agitated throughout the duration of the measurement by a magnetic stirrer, in the container A and the container B.

Nitrogen gas is admitted at a flow rate of between 20 ml / min and 120 ml / min, preferably 80 ml / min in the bottom of the container A. The nitrogen thus causes nitric oxide through the outgoing tubing of the container A into the container B where it will be measured by a device provided for this purpose equipped with an amperometric probe provided with a specific membrane (reference device TBR 1025 World Precision Instruments, ISO-NOP probe). The probe thus delivers a variable current between 10 and 1000 nA proportional to the concentration of nitric oxide in the tested solution, of the order of magnitude of μΜ. The electric current produced by the probe under the influence of nitric oxide is recorded by a suitable device.

Before any measurement, the probe is calibrated by measuring the current produced by the release of known increasing amounts of nitric oxide. It is thus verified that there is a linear relationship between the electric current produced by the probe and the molar concentration of nitric oxide and the correspondence between this electric current and the molar concentration of nitric oxide is determined. For this, 20 ml of the following aqueous solution are placed in the disconnected container B of the container A during the calibration period:

Sulfuric acid (H ₂ S0 ₄ , Sigma-Aldrich reference 32,050-1): 1 ml

Potassium iodide (Kl, Sigma-Aldrich 12636): 3.31 g

Deionized water: 200 ml

Then 50 μΐ, 100 μΐ, 150 μΐ and 200 μΐ of a solution of sodium nitrite (NaN0 ₂ , Sigma-Aldrich number 237213) at a concentration of 360 μΜ are successively added.

The curve corresponding the concentration of nitric oxide in the reaction medium and the electrical intensity produced by the probe expressed in nA is plotted for each addition of the nitric oxide solution.

Evaluation of the neutralizing effect of a molecule to be tested

In a typical measurement, 20 ml of ferrous sulfate solution in vessel A and 20 ml of phosphate buffer pH 7.4 are placed in vessel B. In vessel A, nitrogen gas is admitted for 10 minutes. min, which transits to the container B, to remove the dissolved oxygen present in the solutions of the containers A and B. After these 10 min, the recording of the measurement of nitric oxide is started.

At time T ₀ , the sodium nitrite solution is injected into container A through the flexible polymer stopper by means of a syringe equipped with a needle. The recording of nitric oxide shows a positive evolution that will stabilize briefly after about 3 min, before slowly decreasing to return to baseline in about 20 min. At time Ύ _\ = T ₀ + 4 min, 400 μΐ of hydroethanolic solution is injected at an alcoholic strength of between 0% and 100%, depending on the solubility of the product to be tested, containing a quantity P of product of 4 to 8 mg , depending on the effectiveness of the test product, through the flexible cap of the container B through a syringe equipped with a needle. A control is carried out with 400 μl of solvent in which the product to be tested is dissolved.

The value E of the neutralization effect of nitric oxide by the test product, expressed in μΜ of neutralized nitric oxide per second and per mg of product to be tested, is obtained by the following expression:

(1) E = ^ ^ ^~ ^ ^ ^tem

30P

With: AH _test = difference in concentration of nitric oxide test, expressed in pM of nitric oxide between Ti and T ₂ = T] + 30 sec, obtained with the product solution to be tested.

ΔΗ _1βπ1 = difference in concentration of nitric oxide control, expressed in pM of nitric oxide between ΤΊ and T ₂ = ΤΊ + 30 sec, obtained with the solvent of the product to be tested.

P = quantity of product to be tested in mg.

Example 2. Measurement of the neutralization effect of nitric oxide and effects of various molecules

The test described in Example 1 was used to test the following molecules:

Rosmarinic acid (Aldrich 536954)

P-coumaric acid (Sigma C9008)

(+) Catechin hydrate (Sigma Cl 251)

Caffeic acid (Sigma-Aldrich C0625)

Chlorogenic acid (Sigma-Aldrich C3878)

Quercetin dihydrate (Sigma Q0125)

Swine hemoglobin (Sigma H4131)

Verbascoside (HWI Analytik GMGH 0082-05-80)

Luteolin (Sigma L9283)

After having calibrated the probe for measuring the nitric oxide concentration of the apparatus described in Example 1, in the container A, as described in the example

1 and illustrated in FIG. 1, 20 ml of an aqueous solution of:

Sulfuric acid (H ₂ S0 ₄ , Sigma-Aldrich reference 32,050-1): 1 ml

Ferrous sulphate (FeSO ₄ , 7H ₂ O, Sigma-Aldrich number 215422): 5.5 g

Deionized water: 200 ml

In container B, as described above and FIG. 1, 20 ml of phosphate buffer solution at pH 7.4 are placed.

Nitrogen gas is admitted into the apparatus at a rate of 80 ml / min for 10 minutes, then the recording of the value of the current intensity is started. generated by the nitric oxide detection probe and 0.5 ml of an aqueous solution of sodium nitrite (NaN0 ₂ , Sigma-Aldrich number 237213) is injected (at time T ₀ ) into the vessel A). at a concentration of 0.70%.

At time ΤΊ = T ₀ + 4 min, 400 μΐ of hydroethanolic solution is injected at an alcoholic strength of between 0% and 100%, depending on the solubility of the product to be tested, containing a quantity P of test product expressed in mg, adjusted according to the effectiveness of the tested product.

The values of nitric oxide concentrations at time T ₀ , Ti = T ₀ + 4 min and T ₂ = Ti + 30 sec are noted. The values of E calculated according to formula (1) are given in Table 1 below.

Table 1.

Example 3. Preparation and measurement of the Γο-quinone effect of caffeic acid

Orthoquinone of caffeic acid:

Caffeoyl-quinone can be obtained crystallized, by oxidation of caffeic acid by Po-chloranil in solution in a mixture of ether and tetrahydrofuran (4/1) at -70 ° C, according to a method described in the prior art (20, 21). According to the method for measuring the neutralization effect of nitric oxide as described in Example 1, caffeoylquinone produces an effect E of 24.36 μΜ. ^. Η ^ ¹ .

A high E value is therefore observed for this molecule compared to the effects produced by molecules already known to be able to neutralize nitric oxide.

Example 4. Preparation of plantain extract and evaluation of its nitric oxide neutralization activity.

In order to determine the method of treatment of the plant and the method of extraction leading to the best nitric oxide neutralization efficiency, dry plant and fresh plant extraction was carried out by hydroalcoholic solutions of titers. varying from 10 to 10 ranging from 0% to 100% of ethanol, either by maceration at room temperature for 48 hours, in solvent volumes corresponding to 10 times the weight of the plant (v / w), or by extraction at boiling point. at reflux for 30 min with solvent volumes also corresponding to 10 times the weight of the plant (v / p). Following these preliminary tests, the following method was chosen because it produces the most active extract:

Aerial parts of Plantage lanceolata are harvested and quickly put in freezing at -18 ° C to ensure their conservation. 1 kg of this batch is crushed coarsely and macerated in 10 liters of 60% aqueous ethanol for 48 hours. This alcoholic strength is defined taking into account the water content of the plant, previously determined by desiccation on a representative sample. At the end of the maceration the plant is removed by sieving, and the extractive solution is filtered through a filter of 0.45 μηι of porosity to eliminate most of the germs present. The solution is then concentrated under reduced pressure so as to obtain a total volume of 1 1, completely free of ethanol.

This solution is then dried by lyophilization to obtain a final weight of 43 g.

The effectiveness E of neutralization of nitric oxide is 0.16

EXAMPLE 5 Preparation of an extract of aerial parts of Plantage lanceolata enriched in o-quinones derived from verbascoside

Aerial parts of Plantago lanceolata are harvested and quickly put in freezing at -18 ° C to ensure their conservation. 1 kg of this lot is crushed coarsely and macerated in 10 liters of 60% aqueous ethanol for 48 h. This alcoholic strength is defined taking into account the water content of the plant, previously determined by desiccation on a representative sample. At the end of the maceration, the plant is removed by sieving, and the extractive solution is filtered on a filter of 0.45 μπι of porosity to eliminate most of the germs present. The solution is then concentrated under reduced pressure so as to obtain a total volume of 1 1, completely free of ethanol.

This concentrated solution is then extracted against the current by 5 times 200 ml of ethyl acetate. The organic solutions are combined, dried over anhydrous sodium sulphate, filtered and then evaporated to dryness under reduced pressure. There is thus obtained 1.12 g of dry extract which is taken up in 25 ml of pure ethanol.

This ethanolic solution is subjected to flash chromatography (equipment Grace Reveleris X2) on a 40 g silica gel column (Grace Reveleris Silica Cartridge 40 g) by a dichloromethane-methanol gradient as in Table 2 below:

Table 2

2 ml fractions which are subjected to the nitric oxide neutralization test are collected. The most active fractions which correspond to the elution volumes of 568 ml to 576 ml, which together provide 17.7 mg of dry matter after evaporation of the solvent under reduced pressure, and which has a neutralization efficiency E of the nitric oxide of 0.96

The presence of Γ-quinone of verbascoside is then verified in high-resolution mass spectrometry in tandem, with electrospray ionization in the negative mode, by the presence of a doubly charged ion at m / z 311 corresponding to the quinone in question and exhibiting the characteristic fragmentation of phenylhydantoic heterosides, in this case akin to that of verbascoside. Example 6. Formula for a cream containing a plantain extract, intended to treat skin inflammations (especially dermatitis, eczema, psoriasis) adapted to body use.

A cream having the following weight formula is prepared according to a method and with standard equipment as known from the prior art:

Purified water: 67.28%

Glycerol (Glycerine, AMI Chemistry): 20%

Cetostearyl alcohol (Lanette O, AMI Chemistry): 7%

Palmitostearic acid (Stearin, Stéarinerie Dubois): 3%

Fresh aerial parts extract

of Plantago lanceolata as described in Example 5: 1%

Benzyl alcohol (Geogard 221, Lonza): 0.87%

Sodium cetostearyl sulphate (Lanette E, AMI Chemistry): 0.7%

Dehydroacetic acid (Geogard 221, Lonza): 0.09%

Sodium hydroxide (Sigma-Aldrich): 0.058%

Example 7. Galenic formula of a cream containing a plantain extract, intended to treat skin inflammations (especially dermatitis, eczema, psoriasis) adapted to use on the face.

Purified water: 67.28%

Glycerol (Glycerine, AMI Chemistry): 10%

Cetostearyl alcohol (Lanette O, AMI Chemistry): 7%

Isopropyl myristate (Isopropyl Myristate, Interchemistry): 3%

Fresh aerial parts extract

of Plantago lanceolata as described in Example 5: 1%

Benzyl alcohol (Geogard, Lonza): 0.87%

Sodium cetostearyl sulphate (Lanette E, AMI Chemistry): 0.7%

Dehydroacetic acid (Geogard, Lonza): 0.09%

Sodium hydroxide (Sigma-Aldrich): 0.058%

EXAMPLE 8. Galenic formula of a washing gel containing a plantain extract, intended for cleaning the hair, face and body of persons with skin inflammations (especially dermatitis, eczema, psoriasis)

Mixture of fatty alcohol ethersulfates (Texapon ASV50, AMI Chemistry): 50% Purified water: 34.03% Glycerol (Glycerine, AMI Chemistry): 10%

Sodium chloride (sodium chloride VWR): 4%

Fresh aerial parts extract

of Plantago lanceolata as described in Example 5: 1% Benzyl alcohol (Geogard 221, Lonza): 0.87%

Dehydroacetic acid (Geogard 221, Lonza): 0.09%

Sodium hydroxide (Sigma-Aldrich): 0.01%

Example 9. Galenic formula of a gel for treating skin inflammations (dermatitis, eczema, psoriasis) adapted to body use and on the face.

Purified water: 95.69%

Carbomer (Carbopol 980, Lubrizol): 2%

Fresh aerial parts extract

of Plantago lanceolata as described in Example 5: 1%

Benzyl alcohol (Geogard 221, Lonza): 0.87%

Dehydroacetic acid (Geogard 221, Lonza): 0.09%

Sodium hydroxide (Sigma-Aldrich): 0.35%

EXAMPLE 10 Galenic Formula of a Cream Containing a Plantain Extract for Treating Acne-Related Inflammations

Purified water: 71.04%

Cetostearyl alcohol (Lanette O, AMI Chemistry): 10%

Essential oil of lemon (Myrtea): 7,5%

Sweet orange essential oil (Myrtea): 7.5%

Fresh aerial parts extract

of Plantago lanceolata as described in Example 5: 1%

Sodium cetostearyl sulphate (Lanette E, AMI Chemistry): 2%

Benzyl alcohol (Geogard 221, Lonza): 0.87%

Dehydroacetic acid (Geogard 221, Lonza): 0.09%

Example 11. Cough syrup formula for cough

Purified water: 44.23%

Sucrose: 44.22%

Glycerol (Glycerine, AMI Chemistry): 5%

Fresh aerial parts extract

of Plantago lanceolata as described in Example 5: 4% Natural raspberry flavor: 1.5%

Citric acid monohydrate: 0.75%

Sodium benzoate: 0.3%

Example 12. Galenic Formula of Capsules for Treating the Inflammatory Demonstrations of Osteoarthritis.

For a gelatin capsule # 0 (Capsugel):

Fresh aerial parts extract

of Plantago lanceolata as described in Example 5: 0.350 g

Maltodextrin: 0.045 g

REFERENCES

1. Vascular endothelial cells synthetize nitric oxide from L-arginine. R. M. Palmer, D.S. Ashton, S. Moncada. Nature, Vol. 333, 6174: 664-666.

2. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. R. M. Palmer, A. G. Ferrige, S. Moncada. Nature, Vol. 327, 6122: 524-526.

3. Macrophage Oxidation of L-Arginine to Nitrite and Nitrate: Nitric Oxide Is an Intermediate. M. A. Marietta, P. S. Yoon, R. Iyengar, C. D. Leaf, J. S. Wishnok. Biochemistry, 1988, 27: 8706-8711.

4. Nitric oxide: a cytotoxic activated macrophage effector molecule. J. B. Hibbs, R. R. Taintor, Z. Vavrin, E. M. Rachlin. Biochemical and Biophysical Communications, 1988, Vol. 157, 1: 87-94.

5. Biosynthesis of nitric oxide from L-Arginine. A pathway for the regulation of cell function and communication. S. Moncada, R.M.J. Palmer, E. A. Higgs. Biochemical Pharmacology, 1989, Vol. 38, 11: 1709-1715.

6. Modulation of acute inflammation by endogenous nitric oxide. A. Lalenti, A. Lanaro, S. Moncada, M. Di Rosa. European Journal of Pharmacology, 1992, Vol 211: 177-182.

7. Nitric oxide and its involvement in skin homeostasis and disease - A review. D. Bruch-Gerharz, T. Ruzicka, V. Kolb-Bachofen. Arch. Dermatol. Res., 1998, 290: 643-651.

8. Nitric Oxide Levels in Patients with Psoriasis Treated with Methotrexate. N. S. Tekin, N. Ilter, B. Sancak, Mr. G. Ozden, Mr. A. Gurer. Mediators of Inflammation, 2006, Vol. 2006: 1-5.

9. New markers of disease in atopic dermatitis. A. H. A. Mohsen, H. A. Wahab, E. Allam. Journal of American Science, 2011, Vol. 7 (10): 404-408.

10. Nitric oxide function in the skin. M.-M. Cals-Grierson, A.D. Ormerod. Nitric Oxide, 2004, Vol. 10: 179-193.

11. The role of nitric oxide in sepsis - an overview. K. A. Kirkeb0en, A. Strand. Acta Anaesthesiol. Scand., 1999, Vol. 43: 275-288.

12. Role of Nitric Oxide in Anaphylactic Shock. H. Mitsuhata, R. Shimizu, Mr. M. Yokoyama. Journal of Clinical Immunology, 1995, Vol. 15 (6).

13. Arginine-Based Inhibitors of Nitric Oxide Synthase: Therapeutic Potential and Challenges. J. Viseecek, A. Lojek, G. Valacchi, L. Kubala. Mediators of Inflammation, 2012, Vol. 2012: 1-22. 14. Inhibition of nitric oxide synthase during sepsis: revival because of isoform selectivity, W. Stahl, M. Matejovic, P. Radermacher. Shock, 2010, Vol. 34, (3): 321-322.

15. Our inhibitors for treatment of wrinkles. S. Fujii, E. Lerner. PCT / US2002 / 002292.

16. NO synthase inhibitors and use. Mr. Cals-Grierson. Patent PCT / FR2002 / 002064.

17. Beneficial effect of carboxy-PTIO on hemodynamic and blood gas changes in septic shock dogs. Chieko Mitaka, Yukio Hirata, Kuninori Yokoyama, Takashi Nagura, Yukio Tsunoda and Keisuke Amaha. Crit Care 1997, 1:45.

18. Ruthenium complexes as nitric oxide scavengers: a potential therapeutic approach to nitric oxide-mediated diseases. Fricker SP, Slade E, Powell NA, Vaughan OJ, Henderson GR, Murrer BA, Megson IL, Bisland SK, Flitney FW. Br. J. Pharmacol. 1997, 122 (7): 1441-9.

19. Assessment report on Plantago lanceolata L., folium. Collective work. European Medicines Agency. EMA / HMPC / 437859/2010. Committee on Herbal Medicinal Products (HMPC).

20. The synthesis and isolation of caffeoquinone and caffeoquinone methyl ester. Davies R. Tetrahedron Letters, 1976, 4: 313-314.

21. Oxidation Products of Caffeic Acid as Model Substances for the Antigonadotropic Activity of Plant Extracts. John Mathias, Elans Gerd Gumbinger, and Hilke Winterhoff. Planta Medica, 1990, 56: 14-18.

22. Nitric oxide in arthritis. Daniel Jang and George A. C. Murrell. Free Radical Biology & Medicine, 1998, 24 (9): 1511-1519.

Claims

1. Formula compound

wherein R is selected from the group consisting of:

• -CH = CH-COOR ²

with R ^{2 being} able to represent:

- H, a linear alkyl chain CC ₃ , optionally substituted with one or more functions selected from hydroxyl functions and carboxylic acid, by a benzene ring, a diphenol or a caffeoyl radical;

A saturated or unsaturated C ₆ ring, optionally substituted with one or more functions chosen from hydroxyl and carboxylic acid functions and / or with a caffeoyl radical;

The formula -Y-O-X- (3,4) diphenol (II) with:

. Y is a pyranose ring, substituted with a CH ₂ -OH and at least one hydroxyl function and optionally substituted with a rhamnose;

. X can represent the chain -CH ₂ -C¾- or -CH ₂ -CH (OH) ~;

• -ZOR ³

With:

Z may represent the chain -CH ₂ -CH ₂ -, -CH (OH) -CH ₂ - or -CH ₂ -CH (COOH) -;

R ³ may represent:

. - CO-CH = CH- (3,4) diphenol or

. The formula -W-O-CO-CH =CH- (3,4) diphenol (III) with W is a pyranose ring, substituted with a CH ₂ -OH and at least one hydroxyl function and optionally substituted with a rhamnose;

2. Compound according to claim 1, characterized in that R is selected from the group consisting of:

• -CH = H-COOR ²

with R ^{2 being} able to represent:

- H or

The formula -Y-O-X- (3,4) diphenol (II) with:

. Y represents a pyranose ring, substituted by a CH ₂ -OH and at least one hydroxyl function and substituted by a rhamnose;

. X represents the chain -CH ₂ -CH ₂ -;

• -Z -OR ³

With:

Z represents the chain -CH ₂ -CH ₂ -;

- R ³ represents the formula -W-O-CO-CH = CH- (3,4) diphenol (III) with W represents a pyranose ring, substituted with a CH ₂ -OH and at least one hydroxyl function and substituted by a rhamnose.

3. Compound according to claim 1, characterized in that it has a formula

(la) as

HC = CH-COOR ²

O (la)

with R ² being as defined in claim 1.

4. The compound of claim 1 selected from caffeoyl-quinone, the ^we form the quinone of verbascoside and the ^2nd shape of the quinone of verbascoside.

5. Plantago lanceolata extract enriched with verbascoside quinone.

6. Plantago lanceolata extract enriched in quinone verbascoside, characterized in that it is obtained from the aerial parts of the plant by alcoholic maceration.

7. Compounds according to any one of claims 1 to 4 or extract according to claim 5 or 6, for its use for the prevention and / or treatment of inflammatory diseases and / or disorders.

8. Compounds according to any one of claims 1 to 4 or extract according to claim 5 or 6, for its use for the prevention and / or treatment of diseases and / or inflammatory skin disorders selected from psoriasis, atopic dermatitis, contact dermatitis, skin irritation, contact hypersensitivity reaction, allergic skin manifestations, excessive vasodilatation, rosacea, solar erythema, acne.

9. Compounds according to any one of claims 1 to 4 or extract according to claim 5 or 6, for its use for the prevention and / or treatment of diseases and / or inflammatory joint disorders selected from arthritis, osteoarthritis , rheumatoid arthritis, ankylosing spondylitis, lupus erythematosus, chondritis.

10. Compounds according to any one of claims 1 to 4 or extract according to claim 5 or 6, for its use for the prevention and / or treatment of coughing.

11. Compounds according to any one of claims 1 to 4 or extract according to claim 5 or 6, for its use for the prevention and / or treatment of anaphylactic shock or septic shock.

12. Compounds according to any one of claims 1 to 4 or extract according to claim 5 or 6, for use in combating the intrinsic or extrinsic aging of the skin; to fight against the signs of skin aging, such as photoaging, wrinkles, fine lines, dry or cracked skin; to fight neurogenic skin inflammatory processes; to improve the comfort of sensitive skin; to strengthen the barrier function of the skin; to stimulate the hydration of the skin; to improve the comfort of dry skin; to control sweating; to stimulate lipolysis; to inhibit hair loss.

13. Pharmaceutical or cosmetic composition containing at least one compound of formula (I) as described in one of claims 1 to 4 or an extract according to claim 5 or 6 and a physiologically acceptable vehicle.

14. The composition of claim 13, characterized in that it is intended to be applied to the skin or to be administered orally.

15. Composition according to claim 14, characterized in that it is a cream, a gel, an oil, a lotion, a milk.

16. The composition of claim 14, characterized in that it is a syrup, a lozenge to be sucked, a capsule, a tablet.